CN209162064U - Drop formation system for digital pcr detection - Google Patents

Abstract

The utility model discloses a kind of drop formation systems for digital pcr detection, wherein drop formation system includes: microchannel, has the first opening and the second opening passed in and out for liquid；Rotary drive mechanism is used to that microchannel to be driven to carry out reciprocally swinging；Fluid driving mechanism is used to drive fluids through microchannel；Digital pcr chip comprising chip body with drop storage chamber, is uprightly arranged on chip body and the accommodating cavity being connected to inlet, the leakage fluid dram, the first passage by inlet is connected to drop storage chamber, the second channel for being connected to leakage fluid dram with drop storage chamber that are arranged on chip body the inlet being arranged on chip body；In the accommodating cavity that first opening end of microchannel is inserted into and under the driving of rotary drive mechanism can in accommodating cavity reciprocally swinging.The drop formation system collection droplet formation and it is detected on one, significant more accurate testing result can be obtained.

Description

Drop formation system for digital pcr detection

Technical field

The utility model relates to a kind of drop formation systems for digital pcr detection.

Background technique

Polymerase chain (PCR) reaction technology is modern biology one of the most important instruments, it is widely used in medicine Diagnosis, Personalized medicine, Food Inspection, genetically modified organism detection, Causal Agent Identification, immunoassay, forensic science etc..And Had more than traditional qPCR as the latest generation of round pcr based on the digital pcr (dPCR) that microflow control technique development generates Small reaction volume, faster reaction speed, lower system noise and higher sensitivity.

Drop digital pcr (Droplet Digital PCR) technology is the Water-In-Oil drop technique based on micro-fluidic chip, Single DNA molecules are encapsulated into individual droplet by it by Water-In-Oil structure, using between the inertia realization DNA molecular of oil Mutually isolated, each DNA molecular, which is limited in the droplet of oneself, individually to be expanded, and avoids competing from other sequences. After the amplification for completing DNA molecular under the conditions of suitable temperature, by recording the overall number of droplet and can detecte fluorescence The droplet number of signal can realize the accurate quantification to DNA copy number using Poisson distribution algorithm.

The core technology of submicroliter fluid handling first is that the liquid of microlitre magnitude is divided further into nanoliter even picoliters Micro- reaction system of volume.The major technique branch that micro- reaction system generates is that emulsification microlayer model generates.In recent years, Reported in document a variety of microlayer model generation technique such as membrane emulsifications, spraying emulsion process, micro-fluidic chip method, microchannel injection/ Gunite etc..Recent patent number is the Chinese utility model patent and Publication No. CN104815709A of ZL201410655191.5 Chinese patent application advanced optimized by microchannel generate emulsification microlayer model method.Emulsification these methods of microlayer model exist It is all respectively had some disadvantages in practical application.The method of the Chinese utility model patent of Patent No. ZL201410655191.5 Using interface energy and hydrodynamic shear of the micro liquid when gas liquid film converts, the surface for overcoming liquid to export in microchannel Tension and adhesive force enable the drop for flowing out microchannel nozzle to be successfully detached from microchannel, and are formed greatly in immiscible liquid Small controllable drop.But this method needs microchannel cutting movement above and below liquid level, needs to microchannel relative to liquid level Starting and final position carry out high-precision positioning, it is difficult in Project Realization.Publication No. CN104815709A's The method of Chinese patent application cuts off note by the shearing force of circumference of the microchannel in liquid or the generation of spiral uniform motion The immiscible liquid that enters and form drop, but this method generates the size of drop by various system factors due to microchannel Be affected (such as viscosity, the temperature of environment, movement velocity, motion profile of liquid etc.), and this error of variation It can increase with the quantity for generating drop and accumulate, thus the control difficulty of the volume size homogeneity of high-volume drop formation is very Greatly.

In the prior art, there are also digital pcr chipset drop formations and drop collection to be stored in one, so that drop It can directly be used as detecting after collection, however, the chip of this kind of structure is not easy to stably generate the stabilization of drop or drop Property and homogeneity are poor；Chip detection flux is small, is not able to satisfy high-throughput liquid drop analysis in clinically automation and unit area Requirement；In addition, this kind of chip structure complexity, requirement on machining accuracy are high, it is at high cost.

Summary of the invention

It is existing to solve the purpose of this utility model is to provide a kind of drop formation system for digital pcr detection Drop formation system present in one or more it is insufficient.

In order to achieve the above objectives, a kind of technical solution that the utility model uses is: a kind of liquid for digital pcr detection Generation system is dripped, the drop formation system includes:

Microchannel has the first opening and the second opening passed in and out for liquid；

Rotary drive mechanism is used to that the microchannel to be driven to carry out reciprocally swinging；

Fluid driving mechanism is used to drive fluids through the microchannel；

Digital pcr chip comprising the chip body with drop storage chamber, the feed liquor being arranged on the chip body Mouth, is arranged on the chip body accommodating cavity for being uprightly arranged on the chip body and being connected to the inlet Leakage fluid dram, the first passage for being connected to the inlet with the drop storage chamber store the leakage fluid dram and the drop The second channel of chamber connection, the first passage have the first inner gateway being located inside the chip body, and described second is logical Road has the second inner gateway being located inside chip body；

First opening end of the microchannel is inserted into the accommodating cavity and drives in the rotation Under the driving of mechanism can in the accommodating cavity reciprocally swinging.

Some preferred embodiments according to the present utility model, the accommodating cavity prolong upwards from the upper surface of the chip body It stretches, the inlet is located at the bottom of the accommodating cavity.

Some preferred embodiments according to the present utility model, the accommodating cavity and the chip body, which are integrally formed, to be arranged, or Person, the accommodating cavity are mutually fixedly connected with the chip body.

Some preferred embodiments according to the present utility model, the first passage are connected in end and the drop storage chamber, The second channel is connected in end and the drop storage chamber, and first inner gateway is divided into second inner gateway The different two sides of the drop storage chamber.

According to the utility model, the drop storage chamber has the first communication port being connected to the first passage and institute State the second communication port of second channel connection, the setting of the second communication port described in the first communication port face.

Some preferred embodiments according to the present utility model, the drop storage chamber have arc angling, first connection Mouth is set at the arc angling.

Further, the drop storage chamber is square or rectangle, first communication port and second communication port Be divided into the drop storage chamber it is diagonal on, first inner gateway and second inner gateway are divided into the drop and store The opposite two sides of chamber, first inner gateway and second inner gateway are respectively in end and first communication port, the second company Port connection.

According to the utility model, the first passage, second channel a part or whole part be curve-like.

Some preferred embodiments according to the present utility model, the first passage include at least one straight extended segment and at least One arc extended segment, and be connected to by one end of the straight extended segment with the inlet, at least one described straight extends The inner space of section and at least one arc extended segment constitutes first inner gateway.

Further, first inner gateway by a straight extended segment and an arc extended segment inner space structure It is located at the outside of the drop storage chamber and parallel with a line of the drop storage chamber at, the straight extended segment, it is described The direction of one end of straight extended segment towards the drop storage chamber bends and extends to form the arc extended segment, the arc The end of the separate straight extended segment of extended segment is connected to the drop storage chamber.

As a kind of specific preferred embodiment, the drop storage chamber, the first passage, the second channel constitute in Heart symmetrical structure.

According to the utility model, the inlet is higher than the first passage in the height of vertical direction, and/or, it is described Leakage fluid dram is higher than the second channel in the height of vertical direction.

Preferably, the internal diameter of the inlet is 4mm-8mm, is highly 5mm-15mm；And/or the drop storage chamber Length and width be respectively 5-30mm, be highly 50-1000um.

According to the utility model, the drop formation system further includes the drain being erectly set on the chip body Pipe, the drain pipe are connected to the leakage fluid dram.

Further, the upper surface of the drain pipe from the chip body upwardly extends, with the chip body one Body formed setting is fixedly connected.

Further, the leakage fluid dram is equipped with the negative pressure connector for the outlet mating with negative pressure device.

Some preferred embodiments according to the present utility model, the drop storage chamber, the first passage and described second are led to Road collectively forms a chip unit, and multiple chip units are provided on the chip body.

Some preferred embodiments according to the present utility model, the reciprocally swinging of the microchannel are horizontal hunting.

Some preferred embodiments according to the present utility model have between the first opening and the second opening of the microchannel and hold Product is the liquid storage chamber of 10 μ of μ L~100 L.

According to the utility model, the fluid driving mechanism includes syringe, delivery pipe, the liquid in-out mouth of the syringe By the second open communication of the delivery pipe and the microchannel, the internal diameter of the delivery pipe is less than the microchannel Internal diameter.

Further, the fluid driving mechanism further includes the injector drive group for driving the syringe work Part.

Further embodiment according to the present utility model, the injector drive component include feed screw nut's driving mechanism Or rack pinion driving mechanism.

Further embodiment according to the present utility model, the drop formation system further include described with liquid outlet Fluid reservoir, the liquid outlet of the fluid reservoir, the liquid in-out mouth of the syringe, the delivery pipe one end three changed by threeway It is connected to valve.

According to the utility model, the driving mechanism and the microchannel are mutually detachably connected.

Further, the driving mechanism includes rotating electric machine, rotary shaft and connector, the output end of the rotating electric machine with The rotary shaft is connected, and the connector is fixedly connected on the rotary shaft along the direction perpendicular with the rotating shaft axis On, the microchannel is removably mounted on the connector.

Further embodiment according to the present utility model, the fluid driving mechanism include syringe, delivery pipe, described Connector is tubulose, has the first liquid entrance and second liquid inlet and outlet, one end of the delivery pipe and the institute of internal connection The liquid in-out mouth connection of syringe is stated, the first liquid entrance of the other end and the connector is connected, and the second of the microchannel The second liquid of opening end and the connector, which is imported and exported, to be connected.

Further embodiment according to the present utility model, is provided with multiple connectors in a rotary shaft, and one Multiple delivery pipes are connected on a connector.

Further, the drop formation system further includes withdraw of the needle mechanism, and the withdraw of the needle structure is for making the microchannel It is separated with the connector.

Further embodiment according to the present utility model, the second opening end of the microchannel are set in described Connector an end on, the withdraw of the needle mechanism includes the withdraw of the needle plate that can be slidably arranged on the connector and driving institute The withdraw of the needle plate driving assembly for stating the sliding of withdraw of the needle plate contradicts the microchannel by withdraw of the needle plate sliding, makes microchannel and institute State connector separation.

Further embodiment according to the present utility model, the withdraw of the needle plate driving assembly be feed screw nut's driving structure or Cylinder driving structure.

According to the utility model, the drop formation system further includes pedestal, the rotary drive mechanism can on Lower slider it is arranged on the pedestal, the drop formation system further includes for driving the driving mechanism to slide Vertically move driving mechanism.

According to the utility model, drop formation system further includes the negative pressure device being used cooperatively with the pcr chip, described Negative pressure device generates negative pressure for making in the first passage, drop storage chamber and second channel.

According to the utility model, " the oily phase ", " water phase " have the general sense of this field, are not particularly limited. The density of oily phase is usually less than water phase.

Due to the application of the above technical scheme, the utility model has the advantage that compared with prior art

The utility model provides the new drop formation system of one kind and generates thinking.The drop formation system collection droplet formation Be detected on one, can not only generate the uniform drop of volume in large quantity, and drop can directly be used as examining It surveys, and drop can be realized uniform tiling in drop storage chamber, help to obtain significant more accurate testing result.The liquid Drip the significant advantage that generation system has structure simple, at low cost.The digital pcr detection system of the utility model has detection logical The many advantages such as amount is high, testing result is more acurrate.

Detailed description of the invention

Fig. 1 is the structural schematic diagram for the microlayer model generating means that the utility model uses；

Fig. 2 is the reciprocally swinging schematic diagram for the microlayer model generating means that the utility model uses；

Fig. 3 is that the microlayer model for the microlayer model generating means that the utility model uses generates schematic diagram；

Fig. 4 is situation schematic diagram microchannel and any vibrating motor not being connected in the prior art；

Fig. 5 is feelings microchannel being fixed in the prior art on the vibrating motor that one can produce linear uniform motion Shape schematic diagram；

Microchannel is fixed on one in the microlayer model generating means that Fig. 6 uses for the utility model can be with reciprocally swinging machine Situation schematic diagram on structure；

Fig. 7 is the Study on Relative Factors schematic diagram for influencing microlayer model and generating；

Fig. 8 is the main view of the utility model microlayer model generating means embodiment；

Fig. 9 is the front sectional view of the utility model microlayer model generating means embodiment；

Figure 10 is the left view of the utility model microlayer model generating means embodiment；

Figure 11 is the rearview of the utility model microlayer model generating means embodiment；

Figure 12 is the cross-sectional view at the A-A in Figure 11；

Figure 13 is the partial enlarged view in Figure 12 at C；

Figure 14 is the partial enlarged view in Figure 12 at D；

Figure 15 is the cross-sectional view at the B-B in Figure 12；

Figure 16 is the rearview for the microlayer model generating means another kind embodiment that the utility model uses；

Figure 17 is the front sectional view for the microlayer model generating means another kind embodiment that the utility model uses；

Figure 18 is the main view for the microlayer model generating means another kind embodiment that the utility model uses；

Figure 19 is the left view for the microlayer model generating means another kind embodiment that the utility model uses；

Figure 20 is closed-loop control vibration angle involved in the microlayer model generating means of the utility model use or position Principle of Electric Engine figure；

Attached drawing 21 is the structural schematic diagram of the digital pcr chip system embodiment 1 of the utility model；

Attached drawing 22 is the structural decomposition diagram of the digital pcr chip system embodiment 1 of the utility model；

Attached drawing 23 is the main view of chip substrate in the digital pcr chip of embodiment 1；

Attached drawing 24 be along Figure 23 M-M to schematic cross-sectional view；

Attached drawing 25 is the top view of the chip body of attached drawing 23；

Attached drawing 26 is the bottom view of the chip body of attached drawing 23；

Attached drawing 27 is the structural schematic diagram of sealing cover in the digital pcr system of embodiment 1；

Attached drawing 28 is the structural decomposition diagram one of the digital pcr chip system embodiment 2 of the utility model；

Attached drawing 29 is the structural decomposition diagram two of the digital pcr chip system embodiment 2 of the utility model；

Attached drawing 30 is the axonometric drawing of chip cover board in the digital pcr chip of embodiment 2；

Attached drawing 31 is the main view of the chip cover board of attached drawing 30；

Attached drawing 32 be along Figure 31 N-N to schematic cross-sectional view；

Attached drawing 33 is the bottom view of the chip cover board of attached drawing 30；

Attached drawing 34 is the top view of the chip cover board of attached drawing 30；

Attached drawing 35 is the axonometric drawing of chip substrate in the digital pcr chip of embodiment 2；

Attached drawing 36 is the top view of attached drawing 35；

Attached drawing 37 to attached drawing 39 is to realize showing for drop tiling during drop enters drop storage chamber through first passage It is intended to；

Attached drawing 40 is schematic top plan view when drop realizes tiling in drop storage chamber；

Attached drawing 41,42 is schematic diagram when drop realizes two layers, three layers of tiling in drop storage chamber.

Specific embodiment

The technical solution of the utility model is further elaborated with specific embodiment with reference to the accompanying drawing.

Firstly, in conjunction with the embodiments and attached drawing, the specific knot for the microlayer model generating means that the utility model uses is described in detail Structure and working principle.It should be appreciated that specific embodiment described herein is not used to limit only to explain the utility model The utility model.

The utility model provides the new drop formation system of one kind and generates thinking.The drop formation system collection droplet formation Be detected on one.Drop formation system provided by the utility model it is basic composition include microchannel, rotary drive mechanism and PCR chip, the part other than pcr chip are referred to as drop formation device again in the present invention.The utility model Drop formation system can not only generate the uniform drop of volume in large quantity, and drop can directly be used as examining It surveys.

As shown in Figure 1, which show the microchannel 100 of drop formation system, rotary drive mechanism 200 etc., microchannel 100 have the first opening 110 for exporting the first liquid 130, and rotary drive mechanism 200 is for driving microchannel 100 to do water Flat reciprocally swinging.As shown in Fig. 2, rotary drive mechanism 200 drives microchannel 100 around 221 reciprocally swinging of rotation center, so that micro- First opening 110 of pipeline 100 also generates reciprocally swinging, to generate microlayer model 131 under the liquid level of second liquid 610.

As shown in Figure 1, for the generation microlayer models 131 for enabling the first opening 110 of microchannel 100 to continue, microlayer model Generating means further include fluid driving mechanism 300, which passes through delivery pipe 310 and the second of microchannel 100 Opening 120 is connected.Second opening 120 of microchannel 100 is communicated with the first opening 110, and fluid driving mechanism 300 can pass through Delivery pipe 310 applies stable driving force into microchannel 100, enables 130 steady and continuous of the first liquid in microchannel 100 Flow out from the first opening 110 and generate microlayer model 131.

Drop formation method provided by the utility model is an extremely complex dynamic process, there is several factors influence The volume of drop formation.Principal element has: the surface tension of drop is (between microchannel opening area, the first and second liquid Surface can gap it is related), the adhesive force (being influenced by pipeline openings of sizes and surface properties) between microchannel opening and drop； Shearing force (speed and droplet surface area moved by second liquid viscosity, microchannel determines), centrifugal force (with the quality of drop, The swing radial acceleration of microchannel is related) and tangential inertial force (swing tangential acceleration and drop mass with microchannel It is directly proportional).Centrifugal force is substantially exactly radial inertial force in fact.

As shown in figure 3, the first opening 110 due to microchannel 100 generates rotation under the driving of rotary drive mechanism 200 Movement, so that the microlayer model that the first liquid 130 in microchannel 100 is formed in the nozzle of the first opening 110 of microchannel 100 131 131 tables of speed and microlayer model moved in the nozzle of the first opening 110 of viscosity, microchannel 100 by second liquid 610 The swing of the nozzle of first opening 110 of the quality and microchannel 100 of shearing force and microlayer model 131 that area determines radially adds The relevant centrifugal force of speed and swing tangential acceleration and microlayer model with the nozzle of the first of microchannel 100 the opening 110 Under the collective effect of the directly proportional tangential inertial force of 131 mass so that microlayer model 131 under the liquid level of second liquid 610 take off The nozzle of the first opening 110 from microchannel 100.

Reciprocally swinging mode (referring to Fig. 6-7) and its other party for the microlayer model generating means that the utility model is used below Formula (referring to Fig. 4-5) is compared analysis, to illustrate the utility model peculiar technical effect obtained.It needs to particularly point out , below all analysis be all to carry out stress point using 130 drop of the first liquid of outflow microchannel as independent object Analysis, indicates 130 drop of the first liquid as independent system using dotted line frame in figures 4-7.

Fig. 4 is situation schematic diagram microchannel and any vibrating motor not being connected in the prior art.In the prior art, when Fluid drive apparatus constantly pass through microchannel 100 at the uniform velocity by the first liquid 130 inject second liquid 610 when, drop can slowly not It is disconnected to increase.Because liquid volume is incompressible, then droplet size is also at the uniform velocity to become larger under conditions of at the uniform velocity injecting.Effect There are the surface tension for keeping drop not depart from and adhesive force on drop, in addition there are also downward gravity.When drop rises to When critical size (volume shown in cut-off rule in referring to fig. 4), suffered gravity overcomes surface tension and adhesive force to fall off.Because of drop Microlitre magnitude gravity must be grown to be possible to overcome tension and adhesive force, this method cannot generate by nanoliter characterized by Microlayer model.

Fig. 5 is feelings microchannel being fixed in the prior art on the vibrating motor that one can produce linear uniform motion Shape schematic diagram.As shown in figure 5, in the prior art, when fluid drive apparatus constantly passes through microchannel 100 at the uniform velocity for the first liquid When 130 injection second liquid 610, drop slowly can constantly increase.Because liquid volume is incompressible, then at the uniform velocity injecting Under the conditions of, droplet size is also at the uniform velocity to become larger.The places different from situation shown in Fig. 4 are, while starting linear motor driving Microchannel 100 is at the uniform velocity moved along a straight line to the left.So, the stress condition of drop is as shown in figure 5, because drop and second liquid There is relative motion between 610 by shearing force to the right, the speed and surface area of shearing force and drop are positively correlated.Thus even In fast situation, this power increases as droplet size increases.Gravity is far smaller than shearing force and is ignored.In a certain critical body Product (referring to shown in Fig. 5 cut-off rule), is exactly drop separation micro-pipe road junction when shearing force overcomes surface tension and adhesive force At the time of.Because of the fluctuation of environment and system, this critical size has disturbance up and down, this is to cause droplet size non-uniform Principal element (referring to shown in dotted line a in Fig. 7).Under this uniform motion, error brought by this disturbance is very big (preceding The prior arts such as first patent mentioned by microchannel drop formation technology and background technology part in Fig. 4-5 are stated all to exist This problem).

Fig. 6 can be on rotary drive mechanism for microchannel is fixed on one in the utility model microlayer model generating means Situation schematic diagram.As shown in fig. 6, constantly passing through microchannel 100 at the uniform velocity for the first liquid 130 injection the when fluid drive apparatus When two liquid 610, by high-frequency swing so that the velocity variations of microchannel 100 have a high frequency modulated, so that liquid The suffered resultant force that gets rid of of drop also has a high frequency modulated.It is by shearing force, centrifugal force and tangential inertial force that this, which gets rid of resultant force, Made of merging.When critical point has disturbance, the power that gets rid of of this high frequency variation is enough to break through critical point in a very short period of time Disturbance so that disturbance bring volumetric errors minimize (because drop fluid drive apparatus driving lower volume at the uniform velocity Increase, so meaning that volumetric errors are small less the time required to breaking through critical disturbance).As shown in Figure 7, here it is apparent that same Under critical disturbance, microchannel, which is fixed on one, to be far smaller than with the volumetric errors 1 that the swing on reciprocally swinging motor generates The volumetric errors 2 at the uniform velocity generated.

As shown in Fig. 8, Fig. 9 and Figure 10, as a preferred embodiment, in the utility model drop formation device Rotary drive mechanism 200 includes rotating electric machine 210, rotary shaft 220 and connector 230, the output end and rotary shaft of rotating electric machine 210 220 are connected, and connector 230 is fixedly connected in rotary shaft 220 along the direction with 220 axis perpendicular of rotary shaft, microchannel 100 are mounted on connector 230.Rotating electric machine 210 can drive rotary shaft 220 and the connector 230 to be around the axis of rotary shaft 220 Center rotary oscillation, so as to drive 100 reciprocally swinging of microchannel.Rotation driving in the utility model drop formation device Mechanism 200 can also be using other rotation drive devices, such as oscillating cylinder, rotating electromagnet etc..

In the present embodiment, microchannel 100 is the tubular structure that both ends all have opening, for the ease of installing microchannel 100, connector 230 is also tubulose, and in conjunction with shown in Figure 13, tubular configured joint 230 has the internal third opening 231 and the 4th being connected Opening 232, delivery pipe 310 are connected in third opening 231, and the second opening 120 of microchannel 100 is connected to the 4th opening 232 On.The fluid driving forces that fluid driving mechanism 300 exports can steadily act on micro-pipe by delivery pipe 310 and connector 230 In road 100, the outflow from the first opening 110 of 130 steady and continuous of the first liquid in microchannel 100 is enable to generate microlayer model 131。

Since the utility model microlayer model generating means can be used in field of biological detection, in order to avoid biomaterial Cross contamination, microchannel 100 are usually disposable, to need each used microchannel 100 from connector It is disassembled on 230, in order to improve removal efficiency, the utility model microlayer model generating means further include withdraw of the needle mechanism 400, in conjunction with Shown in Figure 12 and Figure 13, withdraw of the needle mechanism 400 includes withdraw of the needle plate 410 and withdraw of the needle plate driving assembly 420, is offered on withdraw of the needle plate 410 Withdraw of the needle hole 411, withdraw of the needle hole 411 are set in the outside of connector 230, and the second opening 120 of microchannel 100 is set in the 4th opening 232 outside, and it is opposite with withdraw of the needle plate 410, withdraw of the needle plate driving assembly 420 is for driving withdraw of the needle plate 410 to the side of microchannel 100 To movement, when withdraw of the needle plate 410 is resisted against the second opening 120 of microchannel 100, it is applied with to disengage it to microchannel 100 and connects First 230 squeezing action power, withdraw of the needle plate 410, which continues movement, to release microchannel 100 from connector 230, later, withdraw of the needle plate Driving assembly 420 drives withdraw of the needle plate 410 to move to the direction close to delivery pipe 310, can be set with convenient for next microchannel 100 On connector 230.In addition to withdraw of the needle mechanism provided by the present embodiment, microchannel and connector can also be realized using other structures Separation, clamp microchannel for example, by using claw, pulling microchannel from connector pull-up by the movement of driving claw both makes Separation.

As a preferred embodiment, for the ease of mounting and dismounting microchannel 100, the 4th opening of connector 230 232 outside is in up big and down small round table-like, the resistance that reduction microchannel 100 mounts and dismounts.

Specifically, withdraw of the needle plate driving assembly 420 includes withdraw of the needle plate driving motor 421, the first lead screw 422 and the first lead screw spiral shell Mother 423, withdraw of the needle plate driving motor 421 is fixedly mounted in mounting bracket 240, the output end of withdraw of the needle plate driving motor 421 and the One lead screw 422 is connected, and the first feed screw nut 423 is coupled with the first lead screw 422, withdraw of the needle plate 410 and the first feed screw nut 423 are connected.The rotary motion that first feed screw nut 423 and the cooperation of the first lead screw 422 export withdraw of the needle plate driving motor 421 turns Become the first feed screw nut 423 along the linear motion in 422 axial direction of the first lead screw, so as to drive withdraw of the needle plate 410 to carry out Linear motion, it is of course also possible to use the linear drives component of other forms is to drive withdraw of the needle plate 410.Such as cylinder driving.

Such as Figure 18, the withdraw of the needle plate driving assembly 420 of another scheme includes the first cylinder 1421 and the first fixture nut 1423, the first cylinder 1421 is fixedly mounted in mounting bracket 240,1421 piston rod of the first fixture nut 1423 and the first cylinder 1422 front ends are coupled, and withdraw of the needle plate 410 is connected with the first fixture nut 1423.When gas injects the first cylinder 1421, the One cylinder piston rod, 1421 front end is protruding, is axially moved along piston rod.First fixture nut 1423 and the first cylinder 1421 The axial movement that first cylinder 1421 exports is transferred to withdraw of the needle plate 410 by the cooperation of piston rod front end, so as to drive withdraw of the needle plate 410 move along a straight line, and carry out the withdraw of the needle.

Further, rotary drive mechanism 200 further includes mounting bracket 240, rotating electric machine 210 and withdraw of the needle plate driving motor 421 are respectively and fixedly installed in mounting bracket 240, and the both ends of rotary shaft 220 are spindle rotationally arranged in mounting bracket by bearing In 240, the structure of rotary drive mechanism 200 is enable to step up to gather, stablize.

Further, as shown in connection with fig. 9, microlayer model generating means further include longitudinal moving mechanism 500, vertically move machine Structure 500 includes the first mounting plate 510, longitudinal movement driving assembly 520 and longitudinal sliding motion component 530, and mounting bracket 240 passes through vertical It is mounted on the first mounting plate 510 to slide assemblies 530, longitudinal movement driving assembly 520 is for driving 240 edge of mounting bracket Longitudinal sliding motion component 530 slide.Under the action of vertically moving driving assembly 520, mounting bracket 240 can drive rotation to drive Motivation structure 200 moves in the longitudinal direction, i.e., the connector 230 in rotary shaft 220 can move in the longitudinal direction.Pass through control joint 230 It moves in the longitudinal direction, microchannel 100 on connector 230 synchronizing moving in the longitudinal direction can be driven, when needing microchannel 100 When below the first opening insertion second liquid liquid level, microchannel 100 can be driven to transport downwards by control longitudinal moving mechanism 500 Move predetermined altitude；When needing to remove microchannel 100, microchannel can be driven by control longitudinal moving mechanism 500 100 move upwards.Longitudinal moving mechanism 500 also loads microchannel 100 for connector 230 automatically and provides condition, when need will be micro- When pipeline 100 is mounted on connector 230, microchannel 100 can be placed on to 230 lower section of connector, open the second of microchannel 100 Mouth 120 is directed at connectors 230, and starting longitudinal movement driving assembly 520 drives connector 230 to move down, makes the 4th of connector 230 Opening 232 is inserted into the second opening 120 of microchannel 100, then connector 230 is driven to move up reset again.Also, After loading microchannel 100 on connector 230, longitudinal movement driving assembly 520, which can also drive microchannel 100 to move down, makes the Under the liquid level of one opening, 110 insertion second liquid 610, reciprocally swinging is carried out to manufacture microlayer model.

Specifically, in conjunction with shown in Fig. 8 and Fig. 9, longitudinal movement driving assembly 520 includes longitudinal movement driving motor 521, the Two lead screws 522 and the second feed screw nut 523, longitudinal movement driving motor 521 is fixedly mounted on the first mounting plate 510, longitudinal The output end of mobile driving motor is connected with the second lead screw 522, and the second feed screw nut 523 is coupled with the second lead screw 522, Mounting bracket 240 is connected with the second feed screw nut 523.Second feed screw nut 523 and the cooperation of the second lead screw 522 will longitudinal movements The rotary motion that driving motor 521 exports is changed into the second feed screw nut 523 and transports along the straight line in 522 axial direction of the second lead screw It is dynamic, so as to drive mounting bracket 240 to move along a straight line, it is of course also possible to use the linear drives component of other forms To drive mounting bracket 240.Such as rack drives.

Such as Figure 17, the longitudinal movement driving assembly 520 of another scheme includes that the longitudinal movement gear with power interruption braking drives Dynamic motor 1521, first gear 1522 and the first rack gear 1523, longitudinal movement gear drive motor 1521 are fixedly mounted on installation On bracket 240, the output end for vertically moving driving motor is connected with first gear 1522, and the first rack gear 1523 is fixed on first On mounting plate 510, the first rack gear 1523 is coupled with first gear 1522.First gear 1522 and the first rack gear 1523 are matched The rotary motion that exports of gear drive motor 1521 will be vertically moved by, which closing, is changed into longitudinal movement gear drive motor 1521 and the One gear 1522 is along the linear motion in 1523 axial direction of the first rack gear, so as to drive mounting bracket 240 to carry out straight line fortune It is dynamic, it is of course also possible to use the linear drives component of other forms is to drive mounting bracket 240.

As shown in Figure 10, Figure 11 and Figure 12, the fluid driving mechanism 300 in present embodiment includes syringe 350 and note The liquid in-out mouth of emitter driving assembly 320, syringe 350 is connected by delivery pipe 310 with the second opening 120 of microchannel 100 It is logical.The push rod 351 of syringe 350 slides in the cylinder of syringe 350 under the drive of injector drive component 320, pushes Driving liquid therein is entered in microchannel 100 by delivery pipe 310 and connector 230, the first liquid 130 into microchannel 100 Fluid driving forces are provided.Fluid driving mechanism provided by the utility model is not limited to the above embodiment, for example, can be with Using peristaltic pump, pressure-driven pump, pneumatic-driven pump or driven by electroosmosis pump etc..

Further, as shown in figure 12, fluid driving mechanism 300 further includes three-way diverter valve 330 and fluid reservoir, microchannel 100 the second opening 120, the liquid outlet of the liquid in-out mouth of syringe 350 and fluid reservoir and three interfaces of three-way diverter valve 330 It is connected.Three-way diverter valve 330 at least can control fluid driving mechanism 300 and realize following two mode: one, making syringe 350 liquid in-out mouth is connected with the second opening 120 of microchannel 100, under the drive of injector drive component 320, injection Device 350 provides liquid driven power to microchannel 100, for the first liquid in microchannel 100 to be released from the first opening 110, Or the first liquid is drawn into microchannel 100 from the first opening 110；Two, make the liquid in-out mouth and liquid storage of syringe 350 Tank is connected, and under the drive of injector drive component 320, the driving liquid in fluid reservoir is drawn into injection by syringe 350 In the tube body of device 350, or will be in the driving liquid push-in fluid reservoir in syringe 350.

In conjunction with shown in Fig. 8, Fig. 9 and Figure 11, in order to improve the formation efficiency of microlayer model, implementation as one preferred Mode, microchannel 100, connector 230, delivery pipe 310 and syringe 350 are respectively multiple, and multiple connectors 230 are in rotary shaft 220 On be alternatively arranged, multiple microchannels 100 are separately mounted on a connector 230, the both ends of every delivery pipe 310 respectively with one Microchannel 100 second opening is connected with the first interface of three-way diverter valve 330, the liquid in-out mouth of each syringe 350 and The second interface of three-way diverter valve 330 is connected, and the liquid outlet of fluid reservoir is connected with the third interface of three-way diverter valve 330. Multiple microchannels 100 can carry out the work of microlayer model generation simultaneously under the driving of syringe 350 and rotating electric machine 210, and one The microlayer model that a three-way diverter valve 330 may be implemented to control multiple microchannels 100 simultaneously generates state.

As a preferred embodiment, multiple microchannels 100, connector 230, delivery pipe 310 and note can also be corresponded to Multiple three-way diverter valves 330 are arranged in emitter 350, by multiple three-way diverter valves 330 respectively with multiple delivery pipes 310 and multiple injections Device 350 is connected, in this way can be by the way that multiple three-way diverter valve independent controls are realized with the microlayer model to multiple microchannels 100 Generation state is independently controlled.

Further, in conjunction with shown in Figure 12, Figure 14 and 15, fluid driving mechanism 300 further includes mounting blocks 340, Duo Gesan Logical reversal valve 330 and syringe 350 are fixedly mounted on mounting blocks 340, offered in mounting blocks 340 multiple first runners 341, 342, third flow channels 343 of multiple second flow channels and multiple liquid separation runners 344, the both ends of each first runner 341 respectively with A piece delivery pipe 310 is connected with the first interface of a three-way diverter valve 330, the both ends of each second flow channel 342 respectively with The liquid in-out mouth of one syringe 350 and the second interface of a three-way diverter valve 330 are connected, third flow channel 343 and liquid storage Tank is connected with multiple liquid separation runners 344, and each liquid separation runner 344 is connected with the third interface of a three-way diverter valve 330.

Specifically, as shown in connection with fig. 10, injector drive component 320 includes injector drive motor 321, third lead screw 322 and third feed screw nut 323, the output end of injector drive motor 321 be connected with third lead screw 322, third lead screw spiral shell Mother 323 is coupled with third lead screw 322, and the push rods 351 of multiple syringes 350 passes through connector (not shown) and the Three feed screw nuts 323 are connected.Third feed screw nut 323 cooperates with third lead screw 322 exports injector drive motor 321 Rotary motion be changed into third feed screw nut 323 along the linear motion in 322 axial direction of third lead screw, so as to drive note The push rod 351 of emitter 350 moves along a straight line, it is of course also possible to use the linear drives component of other forms is with driving push rod 351.Such as rack drives.

Such as Figure 16, the injector drive component 320 of another scheme includes the syringe gear driving electricity with power interruption braking Machine 1321, second gear 1322 and the second rack gear 1323, output end and 1322 phase of second gear of injector drive motor 1321 Connection, the second rack gear 1323 are connected with the second mounting plate 360, and the second rack gear 1323 is coupled with second gear 1322, more The push rod 351 of a syringe 350 is connected by connector (not shown) with syringe gear drive motor 1321.Second Gear 1322 and the second rack gear 1323, which cooperate, is changed into syringe gear for the rotary motion that injector drive motor 1321 exports Driving motor 1321 and second gear 1322 are along the linear motion in 1323 axial direction of the second rack gear, so as to drive syringe The 351 of 350 move along a straight line, it is of course also possible to use the linear drives component of other forms is with driving push rod 351.

Further, fluid driving mechanism 300 further includes the second mounting plate 360, mounting blocks 340 and injector drive electricity Machine 321 is fixedly mounted on the second mounting plate 360, and the second mounting plate 360 keeps 300 structure of fluid driving mechanism more compact and steady It is fixed.The first mounting plate can also be merged setting with the second mounting plate simultaneously, to save space, such as shown in Figure 19, can will infused Emitter driving assembly 320, which is mounted on, to be integrated on mounting plate 1360, remaining installation does not change with driving method.

As a preferred embodiment, rotating electric machine 210 can use galvanometer motor, galvanometer motor can be provided surely Fixed and high speed reciprocating rotary wobbling action, and the amplitude of oscillation and frequency can be set as desired, greatly improve the utility model The scope of application of microlayer model generating means.Meanwhile withdraw of the needle plate driving motor 421, longitudinal movement driving motor 521, syringe drive Dynamic motor 321 can use stepper motor, and stepper motor and the cooperation of screw-nut structure can accurately control linear motion Stroke improves the degree of automation.

Preferably, rotating electric machine 210 is vibrated using the motor with closed-loop control vibration angle or position by closed-loop control The motor driven rotary drive mechanism 200 of angle or position carries out reciprocally swinging, thus the swing of accurate control microchannel 100 Track, to be further reduced environment and the disturbance of system bring.This method another advantage is that adjustable system Parameter makes critical size that can reach (arrow is signified in such as Fig. 7) in a swing period.This means that each is revolved Transhipment only generates a drop in the dynamic period.Make the change of the droplet size brought by various environmental factors fluctuate in this way Change will not accumulate next cycle.Thus large batch of it can generate drop of uniform size.This point be also other Announcement passes through advantage not available for mechanical movement generation nanoliter/picoliters magnitude emulsion droplet scheme.

Below in conjunction with the application of Figure 16 elaboration closed-loop control vibration angle or the motor of position in the present invention.Closed loop The motor for controlling vibration angle or position includes the components such as infrared position sensor, control circuit and signal processing circuit.At this In utility model, infrared position sensor is installed in the rotary shaft 220 of rotary drive mechanism 200, passes through infrared position sensing Device is its position signal feedback obtained into control circuit, and control circuit is according to PID automation control principle respectively to anti- The position signal of feedback has done ratio, has integrated, processing of differentiating, and the letter of binding site feedforward and speed ring, electric current loop etc. Number processing circuit, absolute position when realizing motor movement accurately controls.Using closed-loop control vibration angle or the electricity of position Machine can be loaded environmental change by complexity to avoid other vibrating motors and cause the change of vibration position, and engineering is conducive to Upper accurate control droplet size and formation speed.

In the present embodiment, having volume between the first opening 110 and the second opening 120 of microchannel 100 is 10 μ L The liquid storage chamber of~100 μ L, the liquid storage chamber can store a certain amount of first liquid, guarantee that the first liquid generates requirement enough Microlayer model, meanwhile, liquid storage chamber can also prevent the first liquid to be inhaled into connector 230, delivery pipe 310 by microchannel 100 In, safeguards system will not be contaminated by the sample.

Preferably, microchannel 100 can be made of nonrigid material, have certain flexibility.Certain flexibility refers to micro-pipe It is certain that under the driving of rotary drive mechanism 200 the motion path of first opening 110 of microchannel 100 can have in road 100 Standing wave phenomena.Using the microchannel made of the material with certain flexibility, liquid level is generated to further reduce Disturbance be more easier, uniformly so that generating drop, while also further reducing liquid crushing phenomenon generated.

In the present embodiment, microchannel 100 is made of the polypropylene material of low-surface-energy；Delivery pipe 310 is by Teflon (Teflon) material is made.

In embodiments, the spout diameter of the first opening 110 of microchannel 100 is 1 μm -250 μm, it is highly preferred that micro- The spout diameter of first opening 110 of pipeline 100 is 10 μm -100 μm.

Next, being described in detail the structure and its work of digital pcr chip in the utility model in conjunction with the accompanying drawings and embodiments Make principle.

Referring to fig. 21 to Figure 22, which show a kind of digital pcr chips, microchannel described in the utility model 100 (also referred to as output rifle needle in the utility model) and with for generating negative pressure and with the negative pressure device of negative pressure rifle needle 50.

The digital pcr chip of this example comprising the chip body 10 with drop storage chamber 1 is arranged in chip body 10 On inlet 4 and leakage fluid dram 5, and the uprightly accommodating cavity 61 that is arranged on chip body 10 and is connected to inlet 4, chip Ontology 10 further includes the first passage 2 for being respectively connected to inlet 4 with drop storage chamber 1, by leakage fluid dram 5 and drop storage chamber 1 The second channel 3 of connection, wherein first passage 2 has the first inner gateway being located inside chip body 10, second channel 3 With the second inner gateway being located inside chip body 10.

In the digital pcr chip, the upper surface of accommodating cavity 61 from chip body 10 is upwardly extended, and inlet 4 is located at accommodating The bottom of chamber 61.In use, first making in drop storage chamber 1, first passage 2, second channel 3 and accommodating cavity 61 filled with oily phase, so Inject water phase into the oily phase of accommodating cavity 61 using output rifle needle 40 afterwards, and make while injection to export rifle needle 40 carry out it is past Physical pendulum is dynamic, so that forming drop in accommodating cavity 61.The density of water phase is typically larger than oily phase, and being formed by drop will be because of itself weight Power is deposited to the bottom of accommodating cavity 61, enters first passage 2 through inlet 4 after, hence into drop storage chamber 1.

Specifically, the length of the accommodating cavity 61 is 2-30mm, width 2-30mm, is highly 20-1000um.The accommodating cavity 61 may be configured as being fixedly connected with chip body 10, can also be integrally formed and be arranged with chip body 10.In the present embodiment, chip The upper surface of ontology 10 has the feed liquor diversion pipe 6 erect extend upwards, and the lumen of the feed liquor diversion pipe 6 is the accommodating on constituting Chamber 61.

In the digital pcr chip, alternate path 3 is also connected in end and drop storage chamber 1.Preferably, the number In pcr chip, preferably, the first access 2 is connected in end and drop storage chamber 1, and the first inner gateway is set to drop and deposits The side of storage chamber 1；The alternate path 3 is also connected in end and drop storage chamber 1, and the second inner gateway is divided into the first inner gateway The different two sides of drop storage chamber 1 can generate negative pressure via leakage fluid dram by the negative pressure rifle needle 50 of negative pressure device, and auxiliary droplet is slow Slowly gradually enter in drop storage chamber 1 from first passage 2.

On the chip body 10, the bottom surface of first passage 2, second channel 3 and drop storage chamber 1 is preferably arranged to be located at same One height and position, inlet 4 are higher than first passage 2 in the height of vertical direction, and leakage fluid dram 5 is higher than the in the height of vertical direction Two channels 3.The internal diameter of inlet 4 is preferably arranged to 4mm -10mm, in fact it is highly preferred that is 5mm -15mm.First passage 2, second is logical The internal diameter in road 3 is respectively 4mm -10mm.The length and width of drop storage chamber 1 is respectively 2-30mm, is highly 20-1000um.

Referring to fig. 2 shown in 6, drop storage chamber 1 has the first communication port 1a and second channel being connected to first passage 2 The two sides side that be divided into drop storage chamber 1 different the second communication port 1b, the first communication port 1a and the second communication port 1b of 3 connections In portion, it is preferred to use first communication port 1a face the second communication port 1b setting；When the cross section of drop storage chamber 1 uses polygon When structure, the first communication port 1a and the second communication port 1b be preferably disposed on one group of polygon it is diagonal on.

In certain embodiments, drop storage chamber 1 has at least one arc angling, and the first communication port 1a is set to above-mentioned arc At shape chamfering, so that first passage 2 is connected on first communication port 1a and connects with drop storage chamber 1, this is more advantageous to liquid Drop realizes tiling movement after entering drop storage chamber 1.The drop storage chamber 1 can be the polygon with circular arc inner chamfer, or Person is round, ellipse.

When the section of drop storage chamber 1 be polygon when, can be used two of them adjacent edge handover position formed it is above-mentioned Arc angling, can also carry out big chamfered wherein and form arc angling in a line, when cutting for drop storage chamber 1 When face is other irregular shapes, preferably it is also required to carry out big chamfered.As a preferred option, the cross of drop storage chamber 1 Angle in section between at least two adjacent edges is rectangular, and the first communication port 1a is set to the right angle.

In the present embodiment, the cross section setting of drop storage chamber 1 is square, the first communication port 1a and the second communication port 1b On one group is diagonal, as shown in figure 26, and other each interior angles are then all made of the setting of circular arc inner chamfer, are conducive to drop in drop Good stability is kept in storage chamber 1.The square side length of 1 cross section of drop storage chamber is 5-30mm, drop storage chamber 1 height is then 50-1000um.When the diameter of drop reduces, the size dimension of the drop storage chamber 1 can also be reduced It is smaller, and on the one hand the setting of height needs to meet the needs of drop tiling, it on the other hand then needs to take into account formula oil and makes full use of The problem of.

Preferably, a part or whole part of first passage 2 and second channel 3 is curve-like.In the present embodiment, referring to figure Shown in 26, first passage 2 includes the liquid input section 2a that one end is connected with inlet 4, and the other end direction from liquid input section 2a Drop storage chamber 1 goes out liquid section 2b in what curvature extended.Liquid input section 2a extends straight section, is located at drop storage chamber 1 Outside and with drop storage a line of 1 chamber it is parallel, the direction of one end towards the drop storage chamber 1 of liquid input section 2a bends and extends Formed arc-shaped extended segment go out liquid section 2b, out the end of the separate liquid input section 2a of liquid section 2b be connected on the first communication port 1a and It is connected with drop storage chamber 1, which is conducive to put down under gravity after drop enters first passage 2 via inlet 4 Quietly enter in drop storage chamber 1.

Second channel 3 and 2 center symmetric setting of first passage.Specifically, second channel 3 includes one end and 5 phase of leakage fluid dram The drain section 3a of the connection and liquid input section 3b extended from the other end of drain section 3a towards drop storage chamber 1 in curvature, should Liquid input section 3b is in the arc gradually to arch upward far from drain section 3a, the end of liquid input section 3b be connected on the second communication port 1b and and liquid Storage chamber 1 is dripped to connect.

On the whole, first passage 2, drop storage chamber 1, second channel 3 constitute centrosymmetric structure.Structure design Realize the steady conveying of drop, it is ensured that the stability of drop.

Chip body 10 is mainly formed by stacking by chip cover board and chip substrate through-thickness, and chip cover board is plate, Open up fluted on chip substrate, the plate and the groove are overlapped mutually compression and form drop storage chamber 1, first passage 2 And second channel 3.In the present embodiment, downward, chip substrate 101 is located at 102 top of chip cover board, chip cover board to slot opening 102 for transparency glass plate, transparent PC plate, transparent acrylic, COP transparent panel or as made of the non-reflective material such as POM, PP it is black The non-reflective plate of color.Glued or ultrasonic wave or the welding of thermocompression bonding technique can be used between chip substrate 101 and chip cover board 102 To sealing, edge between the two need to keep absolute leakproofness.

In another embodiment as shown in Figure 28 to Figure 36, upward, chip substrate 104 is located at chip cover board to slot opening 103 lower sections, specifically, inlet 4 and leakage fluid dram 5 are provided on chip cover board 103, feed liquor diversion pipe 6 and drain diversion pipe 7 Also it is shaped on chip cover board 103；Drop storage chamber 1, first passage 2 and second channel 3 are then arranged in chip substrate 104 On.Above-mentioned chip cover board 103 is all made of plastics with chip substrate 104 and is made, and is welded between the two using thermocompression bonding technique To sealing.

Referring to fig. 21, shown in Figure 22, Figure 23 and Figure 24, the upper surface of chip body 10, that is, chip substrate 101 is upper Surface, which also has to erect upwards, to be extended and the drain diversion pipe 7 that be connected to leakage fluid dram 5, the drain diversion pipe 7 be then mainly used as and The negative pressure connector that negative pressure rifle needle 50 is coupled, the negative pressure rifle needle 50 and drain diversion pipe 7 are coupled to 1 shape of drop storage chamber At negative pressure.In the present embodiment, feed liquor diversion pipe 6 is set to chip substrate 101 with being integrally formed with what drain diversion pipe 7 used On, certainly, in other some embodiments, it can also first be formed by the way of separately machined and pass through ultrasonic bonding again Or the modes such as glue bonding are connected on chip substrate 101.The nozzle of each feed liquor diversion pipe 6 can be by being detachably connected Sealing cover 201 is sealed, this that is, to accommodating cavity 61 realize seal；The nozzle of drain diversion pipe 7 then can be by sealing film 30 It is sealed.

1 to shown in Figure 26 referring to fig. 2, the inside of chip body 10, drop storage chamber 1, first passage 2 and second channel 3 Chip unit is collectively formed, the multiple said chip units being intervally arranged along its length are provided on the chip body 10, it can Multiple groups loading and analysis detection are carried out simultaneously.Correspondingly, inlet 4, leakage fluid dram 5 and accommodating cavity 61 also have multiple groups, feed liquor water conservancy diversion Pipe 6 and drain diversion pipe 7 are also equipped with multiple groups.To be easily manufactured and easy to operate, what is be adapted to multiple feed liquor diversion pipes 6 is all Sealing cover 201, which is wholely set, forms an integral sealing cover 20, sealing film 30 also as a whole component and can Sealing as all 7 nozzles of drain diversion pipe simultaneously.

The utility model additionally provides the detection method using above-mentioned digital pcr chip or digital pcr detection system simultaneously, It includes the loading step that drop is conveyed to drop storage chamber 1, which includes:

Make to be filled with oil in drop storage chamber 1, first passage 2, second channel 3 and the accommodating cavity 61 of digital pcr chip Phase；

Water phase is injected in the oily phase into accommodating cavity 61 using microchannel 100 (i.e. output rifle needle 40) and while injection So that microchannel 100 is carried out reciprocally swinging, makes to form drop in accommodating cavity 61；

The drop is set to be transported to drop storage chamber 1 through inlet 4, first passage 2.

Wherein, before injecting water phase, preferably make drop storage chamber 1, first passage 2, in second channel 3 full of oily phase. It is preferably maintained in inlet 4 before injecting water phase to be in a sealed state with leakage fluid dram 5, and makes pcr chip horizontal rest 5min More than；After initially forming drop, negative pressure device is connected, oil is promoted mutually to be discharged from leakage fluid dram 5 and drop is promoted to flow to liquid Drip storage chamber 1.

Specific detection process carries out in accordance with the following steps: in advance in the drop storage chamber 1 of chip body 10, first passage 2, it filled with oily phase in second channel 3 and accommodating cavity 61, and sealing cover 201 is respectively adopted seals up feed liquor with sealing film 30 and lead The nozzle of flow tube 6 and drain diversion pipe 7.5min or more is stood to chip body 10, sealing cover 201 is opened, drop formation is filled In the accommodating cavity 61 of the output syringe needle insertion feed liquor diversion pipe 6 for the output rifle needle 40 set, so that port (the i.e. micro-pipe of output syringe needle 110) first opening in road 100 injects water phase under the liquid level of oil phase and makes to export syringe needle progress back and forth while injection It swings, makes to form drop in accommodating cavity 61.The drop of generation is deposited in the bottom of accommodating cavity 61 due to the effect of self gravity, And thering is partial drop to fall to first pipe 2 naturally through inlet 4, the intervention of drop at this time, oil phase liquid face height can be in feed liquor It increased in mouth 4, but not influence the stability of drop formation.After completing drop formation, 7 nozzle of drain diversion pipe is punctured Place sealing film 30 (can in addition be arranged cooperation instrument can actuation mechanism punctured to execute), and make connect negative pressure device negative pressure Rifle needle 50 slowly generates negative pressure, and drop can pass slowly first passage 2 by inlet 4 and deposit into drop as pressure is acted on Storage chamber 1, and fan-shaped region is tiled to drop storage region 1, as shown in figure 37 to figure 39, drop formation and preliminary flat at this time Paving process is completed, and can compress chip body 10 by mechanical structure.

It can be compressed in entire 10 upper surface of chip body during compression or several fixed points compress, and pass through spring Etc. structures carry out compensator or trimmer pressure, according to several fixed points compression then need to avoid exciting light irradiation or camera light path region, So as to carry out real-time fluorescence reading, the motion state of drop at any time is observed.

In addition, may be implemented by adjusting the thickness and area of drop storage chamber 1, the volume of drop and population of samples product Accurately one layer, perhaps as shown in figure 41 two layers or as shown in figure 42 three layers, the even more tiling of multilayer.Assuming that sample Product volume is 20 microlitres, and droplet size is 1 nanoliter, and chip area is 16mm × 16mm, and chip thickness is 125~150 microns, It is one layer that 20,000 drops then generated, which can only tile,.And same 20 microlitres of sample volumes, 1 nanoliter of droplet size, if chip area It is adjusted to 11.5mm × 11.5mm, thickness is adjusted to 200~275 microns, then it is 2 layers that 20,000 drops, which can only tile,.

Drop multilayer observation more high-throughput in unit area may be implemented in the tile mode of this multilayer drop.This It is most important to the whole detection flux for the digital pcr equipment for improving image-type detection drop, it solves this kind of equipment and is faced The low bottleneck problem of detection flux.

The above embodiments are only for explaining the technical ideas and features of the present invention, and its object is to allow be familiar with technique Personage can understand the content of the utility model and be implemented, do not limit the protection scope of the present invention, All equivalent change or modifications according to made by the spirit of the present invention essence, should all cover in the protection scope of the utility model It is interior.

Claims (32)

1. a kind of drop formation system for digital pcr detection, it is characterised in that: the drop formation system includes:

Microchannel has the first opening and the second opening passed in and out for liquid；

Rotary drive mechanism is used to that the microchannel to be driven to carry out reciprocally swinging；

Fluid driving mechanism is used to drive fluids through the microchannel；

Digital pcr chip comprising with drop storage chamber chip body, be arranged on the chip body inlet, Uprightly it is arranged on the chip body and the accommodating cavity being connected to the inlet, the drain being arranged on the chip body Mouth, the first passage for being connected to the inlet with the drop storage chamber connect the leakage fluid dram and the drop storage chamber Logical second channel, the first passage have the first inner gateway being located inside the chip body, the second channel tool There is the second inner gateway being located inside chip body；

First opening end of the microchannel is inserted into the accommodating cavity and in the rotary drive mechanism Driving under can in the accommodating cavity reciprocally swinging.

2. drop formation system according to claim 1, it is characterised in that: the accommodating cavity is upper from the chip body Surface upwardly extends, and the inlet is located at the bottom of the accommodating cavity.

3. drop formation system according to claim 1, it is characterised in that: the accommodating cavity and chip body one Molding setting, alternatively, the accommodating cavity is mutually fixedly connected with the chip body.

4. drop formation system according to claim 1, it is characterised in that: the first passage is in end and the drop Storage chamber is connected, and the second channel is connected in end and the drop storage chamber, and first inner gateway and described second Inner gateway is divided into the different two sides of the drop storage chamber.

5. drop formation system according to claim 1, it is characterised in that: the drop storage chamber has and described first First communication port of channel connection, the second communication port for being connected to the second channel, the described in the first communication port face The setting of two communication ports.

6. drop formation system according to claim 5, it is characterised in that: the drop storage chamber has arc angling, First communication port is set at the arc angling.

7. drop formation system according to claim 5 or 6, it is characterised in that: the drop storage chamber be square or Rectangle, first communication port and second communication port be divided into the drop storage chamber it is diagonal on, lead in described first Road and second inner gateway two sides that be divided into the drop storage chamber opposite lead in first inner gateway and described second Road is connected in end with first communication port, the second communication port respectively.

8. drop formation system according to claim 1, it is characterised in that: the part of the first passage, second channel Or generally curve-like.

9. drop formation system according to claim 8, it is characterised in that: the first passage includes at least one straight Extended segment and at least one arc extended segment, and be connected to by one end of the straight extended segment with the inlet, it is described at least The inner space of one straight extended segment and at least one arc extended segment constitutes first inner gateway.

10. drop formation system according to claim 9, it is characterised in that: first inner gateway is prolonged by a straight The inner space for stretching section and arc extended segment is constituted, the straight extended segment be located at the outside of the drop storage chamber and with The a line of the drop storage chamber is parallel, and one end of the straight extended segment is bent simultaneously towards the direction of the drop storage chamber The arc extended segment is extended to form, the end of the separate straight extended segment of the arc extended segment and the drop storage chamber connect It is logical.

11. according to claim 1, drop formation system described in 8,9 or 10, it is characterised in that: the drop storage chamber, described First passage, the second channel constitute centrosymmetric structure.

12. drop formation system according to claim 1, it is characterised in that: height of the inlet in vertical direction Higher than the first passage, and/or, the leakage fluid dram is higher than the second channel in the height of vertical direction.

13. drop formation system according to claim 1, it is characterised in that: the internal diameter of the inlet is 4mm-10mm, Height is 5mm-15mm；And/or the length and width of the drop storage chamber is respectively 2-30mm, is highly 20-2000um.

14. drop formation system according to claim 1, it is characterised in that: the drop formation system further includes straight The drain pipe being on the spot set on the chip body, the drain pipe are connected to the leakage fluid dram.

15. drop formation system according to claim 14, it is characterised in that: the drain pipe is from the chip body Upper surface upwardly extends, and is integrally formed setting with the chip body or is fixedly connected.

16. according to claim 1 or drop formation system described in 14, it is characterised in that: the leakage fluid dram be equipped with for The negative pressure connector of the outlet mating of negative pressure device.

17. drop formation system according to claim 1, it is characterised in that: the drop storage chamber, the first passage And the second channel collectively forms a chip unit, and multiple chip units are provided on the chip body.

18. drop formation system according to claim 1, which is characterized in that the reciprocally swinging of the microchannel is water Flat pendulum is dynamic.

19. drop formation system according to claim 1, which is characterized in that the first opening of the microchannel and second The liquid storage chamber for being 10 μ of μ L~100 L with volume between opening.

20. drop formation system according to claim 1, it is characterised in that: the fluid driving mechanism include syringe, Delivery pipe, the liquid in-out mouth of the syringe are described defeated by the second open communication of the delivery pipe and the microchannel The internal diameter for sending the internal diameter of pipe to be less than the microchannel.

21. drop formation system according to claim 20, it is characterised in that: the fluid driving mechanism further includes being used for Drive the injector drive component of the syringe work.

22. drop formation system according to claim 21, it is characterised in that: the injector drive component includes screw rod Nut driving mechanism or rack pinion driving mechanism.

23. drop formation system according to claim 20, it is characterised in that: the drop formation system further includes described Fluid reservoir with liquid outlet, the liquid outlet of the fluid reservoir, the liquid in-out mouth of the syringe, the delivery pipe one end three Person is connected by three-way diverter valve.

24. drop formation system according to claim 1, it is characterised in that: the driving mechanism and the microchannel It is mutually detachably connected.

25. drop formation system according to claim 24, it is characterised in that: the driving mechanism include rotating electric machine, Rotary shaft and connector, the output end of the rotating electric machine are connected with the rotary shaft, the connector along with the rotary shaft The direction of axis perpendicular is fixedly connected in the rotary shaft, and the microchannel is removably mounted on the connector.

26. drop formation system according to claim 25, which is characterized in that the fluid driving mechanism includes injection Device, delivery pipe, the connector are tubulose, have the first liquid entrance and second liquid inlet and outlet of internal connection, described defeated One end of pipe is sent to be connected to the liquid in-out mouth of the syringe, the first liquid entrance of the other end and the connector is connected, institute The second liquid inlet and outlet of the second opening end and the connector of stating microchannel are connected.

27. drop formation system according to claim 26, which is characterized in that be provided in a rotary shaft multiple The connector is connected with multiple delivery pipes on one connector.

28. the drop formation system according to claim 25 or 26, which is characterized in that the drop formation system further includes Withdraw of the needle mechanism, the withdraw of the needle structure is for separating the microchannel with the connector.

29. drop formation system according to claim 28, which is characterized in where the second opening of the microchannel End cap is located on an end of the connector, and the withdraw of the needle mechanism includes that can be slidably arranged in moving back on the connector Needle plate and the withdraw of the needle plate driving assembly for driving the withdraw of the needle plate sliding contradict the microchannel by withdraw of the needle plate sliding, Separate microchannel with the connector.

30. drop formation system according to claim 29, it is characterised in that: the withdraw of the needle plate driving assembly is screw rod spiral shell Female driving structure or cylinder driving structure.

31. drop formation system according to claim 1, which is characterized in that the drop formation system further includes base Frame, the rotary drive mechanism, which can slide up and down to, to be arranged on the pedestal, and the drop formation system is also wrapped Include the longitudinal movement driving mechanism for driving the driving mechanism sliding.

32. drop formation system according to claim 1, which is characterized in that drop formation system further includes and the PCR The negative pressure device that chip is used cooperatively, the negative pressure device is for making the first passage, drop storage chamber and second channel Interior generation negative pressure.